The volatile compounds for the different tea samples were detected using a GC-IMS instrument (Flavourspec ® , G.A.S, Dortmund, Germany) according to the method described by Yang et al. [14] (link) and modified slightly. Briefly, 0.3 g of tea samples, ground with a mortar and pestle, were placed into a 50 mL centrifuge tube and 30 mL of boiling water and 80 µg of 2-octanol were added. The samples were heated for 5 min at 70 • C. The extraction was then cooled and vigorously mixed. Then, 5 mL of the supernatants was put into 20 mL sample bottles and subsequently incubated at 70 • C for 20 min at an agitation speed of 500 rpm. Then, 500 µL of the headspace was automatically injected into the inlet via a heated syringe at 65 • C. The chromatographic separation was performed on an MXT-5 capillary column (15 m × 0.53 mm × 1 µm, Restek, Beijing, China). High-purity nitrogen was employed as a carrier gas using the following programmed flow: increased 2 mL/min for 2 min, reaching 100 mL/min within 18 min, and held for 10 min. The drift tube was 98.0 mm long, with a drift gas flow rate of 150 mL/min. The temperatures of the column and drift tube were kept at 60 • C and 45 • C, respectively. The procedure was repeated 3 times for every sample.
Mxt 5 capillary column
Manufactured by Restek
Sourced in United States
The Restek MXT-5 capillary column is a fused silica column with a 5% diphenyl - 95% dimethyl polysiloxane stationary phase. It is designed for the separation of a wide range of organic compounds, including hydrocarbons, halogenated compounds, and oxygenated compounds.
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4 protocols using mxt 5 capillary column
1
Headspace GC-IMS Analysis of Tea Volatiles
2
Volatile Organic Compound Analysis via GC-IMS
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The VOCs in each sample group were detected using the GC-IMS flavor analyzer (FlavourSpec ®, G.A.S. Dortmund, Germany). The procedure was as follows: The sample (2 g) was ground at a low temperature and placed in a 20 mL headspace vial, incubated at 60 °C for 20 min, and then the 500 μL headspace sample was automatically injected into the injector at 85 °C through a heated syringe (non-shunt mode). The sample was transferred into an MXT-5 capillary column (15 m × 0.53 mm, film thickness 1 μm) (Restek, Bellefonte, PA, USA) through high-purity nitrogen (99.99%) and introduced into an ionization chamber after elution at 60 °C (isothermal mode). The sample was later scanned in the drift tube, and each spectrum was scanned 12 times. The VOCs were identified by comparing their retention index and drift time with the standards in the GC-IMS library. The relative quantification of VOCs was based on the peak signal intensity. The fingerprint of VOCs was constructed using GalleryPlot (FlavourSpec ®, G.A.S. Dortmund, Germany) supported by a GC-IMS instrument.
3
Headspace GC-IMS Analysis of VOCs in Fermented Mandarin Fish
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The VOCs in traditional fermented mandarin fish were identified using a FlavourSpec GC-IMS (G.A.S., Dortmund, Germany) equipped with a syringe and an autosampler unit for headspace analysis. Briefly, 2 g of homogenized sample was transferred into a 20-ml headspace vial, and the vial was sealed using a magnetic cap with a silicone septum (Chen et al., 2021 (link)). Next, 500 µl of the headspace volume was injected automatically into the heating injector with a heated syringe at 85°C, and the incubation rotation speed was maintained at 500 rpm. Nitrogen (99.99% pure) was used as a carrier gas to transport the sample into a MXT-5 capillary column (15 m × 0.53 mm ID) (Restek Corp., Bellefonte, PA, United States ) for separation at a flow rate of 150 ml/min. The column temperature was maintained at 60°C, and the IMS temperature was 45°C. The carrier gas flow rate was set to 2 ml/min for 2 min and subsequently increased to 100 ml/min for 18 min (Song et al., 2021 (link)). The VOCs were identified by comparing their cation retention index and drift time with information in the FlavourSpec GC-IMS library and NIST 2014 databases (G.A.S, Dortmund, Germany).
4
GC-MS Analysis of Organic Compounds
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GC-MS was performed with an Agilent 6890 Plus gas chromatograph interfaced with a single-quadrupole Agilent 5975C MSD (Agilent Technologies; Palo Alto, CA, USA). Each sample (2 μL) was injected in split mode (10:1) at 300 °C and separated through a MXT-5 capillary column (15 m × 0.25 mm I.D., 0.25 μm film thickness, Silcosteel-treated stainless steel; Restek; Bellefonte, PA, USA). The oven temperature was initially held at 265 °C for 5 min, then increased to 280 °C at a rate of 2 °C/min, and finally increased to 380 °C at 5 °C/min (held for 3 min). The carrier gas was ultra-high-purity helium at a column head pressure of 89.6 kPa (13 psi; column flow: 1.1 mL/min at an oven temperature of 265 °C).
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